Project Summary/Abstract: POP has become a significant medical burden. In the United States, almost one in four women suffer from one or more symptoms of POP. In this disorder, pelvic organs protrude through the vaginal inlet due to pelvic floor dysfunction. While pelvic floor muscle plays an important role in visceral support, deficiencies in the strength of endopelvic fascia have shown to cause prolapse even without muscle damage. Collagen is the most abundant constituent of endopelvic fascia. With elastin and smooth muscle cells (SMCs), it forms a suspensory system to support the pelvic organs. Evidence suggests that individuals with prolapse have abnormal quantity and function of fibroblast cells, which maintain and remodel the collagen matrix. Consistent with reports in literature, we found reduced collagen content, increased collagen types I (COLI) to III (COLIII) ratio and increased MMP expression in vaginal wall connective tissues of POP patients leading to a loose and fragile fiber network, reduced collagen-smooth muscle integration and reduced load-bearing mechanical capacity. We hypothesize that by transforming the extracted cells from POP patients in vitro and reviving their matrix protein productivity, the application of these transformed cells in vivo can restore the biomechanics of pelvic floor connective tissues. The objective of the proposal is to attest the functionality of the transformed fibroblast cells in decellularized matrices of patient tissues, and to evaluate the feasibility of transplanting the transformed fibroblasts to restore the biological function of vaginal wall connective tissues in the POP mouse model. The proposed research has a great translational potential as the use of autologous fibroblast cells and the choice of local cell administration are simple, safe and direct, making the procedure affordable for most patients.